Screening And Identification Of STIL Gene In Nasopharyngeal Carcinoma And Its Function In CNE-2Z Cells

Nasopharyngeal carcinoma is one of the high-grade malignant tumors in China and ranked first in the Otorhinolaryngologic neoplasms.In southern China and Southeast Asia,the incidence of nasopharyngeal cancer is particularly high.The etiological studies of nasopharyngeal carcinoma have shown: Genetic factors,Epstein-Barr virus infection,environmental factors and dietary habits are closely related to the occurrence of nasopharyngeal carcinoma.At present,the treatment of nasopharyngeal carcinoma is based on radiotherapy,due to its location and high sensitivity to radiation.With the continuous improvement of treatment methods,molecular targeted therapy has become a new approach to treat nasopharyngeal cancer.The molecular targeted therapy combined with radiotherapy and chemotherapy has improved the therapeutic effect.The occurrence of nasopharyngeal carcinoma is a multi-stage,multi-gene and multi-step process,and finding key targets has become a hot topic in the research of targeted therapy for nasopharyngeal carcinoma.STIL is an essential component of centrioles replication,and its expression level is closely related to the number of centrioles.The abnormality of the centrosome will lead to the heterodiploidization of chromosomes in the division.The common features of tumors include centrosome abnormalities and chromosomal instability.The STIL gene may play an important role in the development of nasopharyngeal carcinoma.Studies indicate that STIL is only expressed in proliferating cells and is associated with poor metastasis and poor prognosis in cancer cells.Many studies have shown that the STIL gene is involved in the development of various tumors,such as ovarian cancer,pancreatic cancer,lung cancer,and leukemia.Knockdown of STIL gene can effectively inhibit the proliferation of cancer cells.However,there has been no report on the relationship between STIL gene and nasopharyngeal carcinoma.In this issue,the mRNA microarray was used to analyze nasopharyngeal carcinoma and its adjacent tissues.Genes significantly up-regulated in cancer tissues were selected,and 34 genes were initially screened out using bioinformatics combined with literature screening.Based on these,qPCR assay and HCS cell proliferation screening method was used to rescreen the genes that were positive for the proliferation inhibition of nasopharyngeal carcinoma CNE-2Z cells,and the expression of the rescreened genes in the clinical sample tissues was also performed by qPCR.Finally,target gene was confirmed for subsequent studies.CNE-2Z cells was infected with the lentivirus vectors of target gene RNAi to observe its effect on cell biological behavior.The results showed:1)qPCR detection was performed in CNE-2Z cells.Thirty-two of the 34 genes involved in nasopharyngeal carcinoma were highly expressed;2)Selecting the top 20 genes in the 32 genes for HCS cell proliferation screening.On the 5th day,the Ctrl group cell count fold value is compared with the experimental group cell count fold value as the Fold change value.When Fold change ? 2.0,cell proliferation was significantly slower in the experimental group than in the Ctrl group,and the KLHL42 and STIL genes were selected.3)qPCR was used to detect the expression of KLHL42 and STIL genes in nasopharyngeal carcinoma tissues and their adjacent tissues.KLHL42 was up-regulated in 4 pairs of strong reference and 4 pairs of weak reference samples.STIL was up-regulated in 4 pairs of strong reference and 3 pairs of weak reference samples.With reference to the previous results of the m RNA chips,the STIL gene was determined for downstream experiments;4)Successfully constructed RNA interference lentivirus vector targeting STIL;5)After infection of nasopharyngeal carcinoma cells with lentiviral plasmid,the mRNA and protein expression levels of STIL gene in CNE-2Z cells were significantly inhibited,while inhibiting cell proliferation and inducing apoptosis,and affecting the cell cycle.In conclusion,this study confirms that STIL gene can affect CNE-2Z cells,indicating that STIL gene is expected to be a target for the treatment of nasopharyngeal carcinoma and provide a new target for the treatment of nasopharyngeal carcinoma.